Time in minutes



Dec. 5, 1967 U. TEOTINO ET P -PHENYLPHENACYL DERIVATIVES OF I -HYOSCYAMINE AND a-( TROPYLATROPINE Filed Aug. 17, 1966 PULSE RATE IZO IIO

GASTRIC JUICE TOTAL VOLUME IN ML. N (,0 U1 0 2 SheetsSheet l TIME INHOURS AFTER FEEDING FIG. I. 1

CONTROLS T REAT E D INVENTORS UBERTO TEOTINO DAVIDE DELLA BELLA ATTORNEYS.

Dec. 5, 1967 u. TEOTINO ET AL 3,356,682

P-PHE-NYL-PHENACYL DERIVATIVES OF E 'HYOSCYAMINE ANDo/ 1 TROF'ILATROPINEFiled Aug. l7, 1966 2 Sheets-Sheet 2 CONTROLS z CONTROLS o---o TREATED QTREATED 8D a g I w L 400 Z 300 Q I a; o a w I00 1 I I I ,9 O o I 2 3 4TIME IN HOURS AFTER FEEDING O 0 I 2 3 4 TIME IN HOURS AFTER FEEDING rREPLICATES w 90- ISA INE SOLUTIONI I 1 I I I I l I I I I5 0 I5 30 6O 7590 I05 I20 I35 I TIME IN MINUTES FIG. 6. IATROPINE SULFATEI I l I I l 457 5 90 I05 I20 I35 I50 TIME IN MINUTES IOO PULSE R TE IOO FIG];

PULSE RATE I I I I5 6O 75 I05 I20 I35 I50 TIME IN MINUTES INVENTORS wUBERTO TEOTINO BY DAVIDE DELLA BELLA ya-hl fwf w ATTORNEYS.

(III- United States Patent 3,356,682 p-PHENYLPHENACYL DERIVATIVES 0Fl-HY- GSCYAMINE AND dl-TROPYLATROPINE UhertoTeotino and Davidi DeliaBella, Milan, Italy, assignors to Whitefin Holding S.A., Lugano,Switzerland Filed Aug. 17, 1966, Ser. No. 572,983 4 Claims. (ill.26l)292) ABSTRACT OF THE DISCLOSURE New p-phenylphenacyl derivatives ofl-hyoscyamine and dl-tropylatropine having the formula:

This" application is a continuation-in-pa rt of copending applicationSer. No. 406,893, filed on Oct. 27, 1964, now abandoned. i

This invention relates to new tropyl tropate derivatives possessingvaluable therapeutic properties and to the preparation thereof, as wellas to therapeuti compositions comprised thereof.

- It is known that atropine is a classical powerful antispasmodic andpossesses gastric antisecretory activity. However, its great toxicityand undesirable systemic side effects severely limit its therapeuticapplications.

. Up to this time, several permutations of the molecular structure ofatropine have been carried out in order to prepare a drug endowed withth same degree of antispasmodic and gastric antisecretory activity asatropine but at the same time devoid of its known side effects such asdryness of the mouth, retention of urine, diminution or cessation ofperspiration, reduction of other body secretions, dilation of pupils anddisturbance of heart rhythm. Although some useful synthetic compoundshave 7 resulted from this research, there is wide recognition that sucha drug has not yet been found (A. Burger, Medicinal Chemistry, page 463,Interscience Publishers Inc., New r York, NY. (1960).

We have now found a number of tropyl tropate ea-j rivatives whichpossess a strong antispastic action and gastric antisecretory activitybut no undesirable effects,

clinically useful.

-O-CO- LH-CH OH wherein X is halogen.

It will be apparent to those skilled in the art that, referring only tothe stereo-isomeric forms of tropic acid, the tropyl tropate used asstarting material in the practice of this invention, as well as thequaternary ammonium compounds of the foregoing structural formulaobtained as products therefrom, can exist in three stereoisomeric forms.i

The present invention is concerned with the derivatives of all three.stereo-isomeric forms, namely dl-tropyl tropate (atropine), l-tropyltropate (l-hyoscyamine) and d-tropyl tropate (d-hyoscyamine). 7

Among the tropyl tropate derivatives of this invention, p-phenylphenacyl1 hyoscyaminium bromide is preferred in view of its strong andlong-lasting antispasmodic and gastric antisecretory properties.

Its pharmacological and clinical features may be summarized asfollows:

(a) Its parasympatholytic activity is mainly exerted at the level ofperipheral ganglionic synapses on which the compound is three times asactive as hexamethonium and ten times as'tetraethylammonium. On theother hand, it is about twenty times less eflective than hexamethoniumon ortho-sympathetic ganglionic. synapses.

Since atropine, by contrast, acts with equal potency both at the levelof peripheral ganglionic synapses and on ortho-s'ympathetic ganglionicsynapses, it is immediately evident from these tests how more selectiveis the action 'of the newly discovered compounds of this invention' i(b) It counteracts effectively both motor and secretory activity of thegastrointestinal tract elicited by either peripheral stimulation of thevagus nerve or administrationof acetyl-B-methyl choline. (Salivarysecretion and pupillary muscles are only slightly affected.) In order tomore clearly show this most important prop- "erty ofthe claimedproducts, we report hereinafter some of the many pharmacological testsperformed to this effect inafter be represented by the symbol (K).

The qualitative and quantitative changes induced by the compound (K) ongastric secretion may be deduced from the following Table 1.

TABLE 1 Total acidity (1101) Volume of gastric Total peptic power juice(ml) (PU) Time (l1r.) Meq./ml. Total Meq.

Controls Treated Controls Treated Controls Treated Controls Treated 0. 9l. 2 O. 061 0. 056 0. 054 0. 067 l2. 9 l. 7 O. 122 0. 066 l. 57 0. 11302 266 10.7 2. 8 0. 130 0. 063 1.39 0. 17 248 252 7. 6 3. 4 0. 1020.077 O. 78 0. 26 72 6.5 2. 2 0. 0.086 0.65 0. 18 47 39 The data ofTable l are the mean of 3 values for controls, and the mean of twovalues for experiments on the treated animal. The experiments werecarried out on a male dog, weighing 23 kg., which, four months before,

had been provided by surgical operation with a Pavlov found to elicit amarked inhibition on volume of gastric 5 juice, as well as on HClsecretion, while secretion of pepsin was practically unaffected.

The above and other experimental results are graphical- 1y presented inthe attached drawings wherein:

FIGURE 1 is a graph showing the effect of (K) on the volume of gastricjuice from the Pavlov pouch in a conscious dog after a meat meal as afunction of time in hours after the meal;

FIGURE 2 is a graph similar to FIGURE 1 but showing the effect of (K) onthe total HCl secretion of the FIGURE 3 is a graph similar to FIGURE 1but showing the total peptic power, expressed in pepsin units, in thegastric juice collected after the meat meal from a dog treated with (K)as compared with an untreated dog (control);

FIGURE 4 is a graph comparing the heart frequency of a 21 kg. male do asdetermined by electrocardiograph (ECG), when injected with compound (K)as compared with the use of atropine;

FIGURES 5-7 are graphs of pulse rate at time intervals before and afterthe. injection of saline solution atropine sulfate and (K),respectively, the doses of atropine sulfate and of (K) being varied,equivalent symbols being used for doses of equivalent primary effect.

The very good spasmolytic activity of the products of the invention maybe evaluated from the following table where the antagonistic activityexherted by the product (K) towards different spasmogenic agents on invitro? isolated organs is expressed as percent inhibition of theservation (that is with both the investigator and the patient unaware ofthe identity of the injected product), of 23 patients treatedintravenously with 10 mg. of product (K) 19 experienced very good relieffrom the pain.

(c) Blood pressure, cardiac rhythm and ECG (electrocardiographic)tracing appeared unaltered in both cat and dog following administrationof doses of the derivative up to twenty to forty times the minimumeffective dose.

In order to show how surprisingly lower the side 0 effects of thecompounds of the invention, and in particular of the product (K), are ascompared to those of atropine, we have reported in FIGURE 4 the heartfrequency as recorded by ECGof a male dog, weighing 21 kg., injected onetime with atropine and the other with product (K). The drugs wereinjected through a needle, inserted into the saphenous vein about onehour before administration and connected by a polythene tube to asyringe. The drugs were used at the following doses:

G./kg.

Atropine sulfate 100 Product (K) 100 It is evident how intense and longlasting is the tachycardia caused by atropine, while the effect causedby the product (K), besides being very weak, is promptly reversible.

(d) Parasympatholytic activity of the derivative, investigated byrecording bradycardia and blood pressure fall induced by stimulation ofthe peripheral end of the sectioned right vagus in both cat and dog,proved to be very long-lasting: 4 to 5 hours following mg./kg. dosesintravenously. In other animal species such as rodents, both intensityand duration of action appeared to be less significant.

Results in man were comparable to those obtained in cat and dog: 10-l2mg. intravenous doses, 5 to 6 times as high as those which wereeffectively antispasmodic and gastrically antiscretory, did not causexerostomia, mydriasis or heart rhythm disturbance.

This is, for example, demonstrated by the pulse rate (cardicaccelerating activity), measured by palpation of theleft radial artery,reported in FIGURES 5-7. While atropine sulfate induces remarkablealterations of the pulse rate, product (K) practically does not affectthe spasm, normal rate. The experiments were performed on groups.

TABLE 2 Concen- Isolated organ Species Stimulating agent tration Concen-Percent tration inhibit Ileum Guinea pig. Aeetyleholine eh1oride 1X10"1X10 95 Tetramethylammonium iodlde.... 5X10- 1X10- Nicotine tartrate 2.5X10- 5X10 50 5-hydroxytryptamine 1 10 1X10; Bradykinin 2.5x10- ggg,Duodenum Rabbit..- Spontaneous aetlvity 1X1tl- Acetylcholine chloride5X10" IXlO- 85 Vagus n. stomach. Rat Electrical stimulation 3g UterusRat Spontaneous activity 3g Acetylchloine chloride 1 10 2. 5X10" 80 lTone reduction.

2 Estrus phase.

These results have been confirmed in clinical tests, wherein, on 27patients affected by colicky pains due to different causes, treated byintravenous injections with 10 mg. of product (K) under single-blindobservation (that is with only the investigator aware of the productinjected, while it was unknown to the patent) 24 patients experiencedvery good or good relief from the pain. In another set of testsperformed under double-blind obof 9 subjects. The values reported in thefigures are the mean pulse rate as measured on groups of 9 patients in12 experiments. The time 0 corresponds to the moment the saline solutionor the drug is injected, while the negative times indicate testsperformed before any treatment of the patient in order to state acomparison basis, and the base line in each graph represents the mean ofall pre-treatment values.

The doses of atropine sulfate and of product (K) injected are not equalin absolute amount, but have been calculated on the basis of beingequivalent in that they cause the same primary elfect.

Also, for the absence of xerostomia induced by the drugs of theinvention, the following tests (some of the many clinical testsperformed to this end) are indicative of the difference existing betweenatropine, which causes a remarkable diminution of salivary secretion,and the product (K) of the invention, which at the therapeutic doses notonly does not cause xerostomia, but on the contrary, seems to slightlyfavor salivary secretion.

The salivary secretion has been estimated with a method similar to thatused by Brownlee et al. (Clin. Pharm. & Ther. 6,177, 1965). The resultsare reported in Table 3.

product thus obtained with an O-acyl-tropic-acid halide and deacylatingthe resulting compound.

The compounds of the present invention and the processes for preparingthem are illustrated by the following examples.

Example I TABLE 3.-SALIVARY SECRETION IN ML. BEFORE AND AT THREE TIMEINTERVALS AFTER I.V. TREATMENT OF THE 9 TEST SUBJECTS Times at whichsalivary secretion tests were performed:

Before: Min. 30, 15 After:

Min. 5, 10, 15, 20, 25, (lst interval) Min. 45, 60, 75, 9 (2d interval)Min. 105, 120, 135, 150 (3d interval) After Treatment Before (BasalValues) Grand means of the difierences from the basal values Treatmentsand Doses Grand means of the means of 2 tests 1st interval: 2d interval:3d interval" per subject 30 min. 1 hr. 1 hr.

(6 tests per (4 tests per (4 tests per subject) subject) subject) SalineSolution 1 1. 03:0. 2 0. 4 0. 3 O. 2 1. 2i0. 3 O. 4 0. 3 0. 2 1. 33:0. 20. 3 0 O. 1

Atropine Sulfate, m

0.25 1.2i0.3 0.5 0.3 0.3 1. 210. 2 0. s 0. 5 0. 1 1.55:0. 2 l. 2 -l. 00. 8

1 3 replicates.

From the above reported data it is evident that product (K) up to 10 mg.intravenous doses (which are at the maximum level of the therapeuticdoses) does not possess atropine-like properties to any significantextent.

In summary, the products of the invention, and in particularpphenylphenacyl-hyoscyaminium bromide (product K), are characterized bya very high (and in our experience unique) ratio of inhibitory activityon the muscarinic receptor and the orthosympathetic ganglia and thusprove to have very good antispastic and antisecretory effects whilebeing practically completely free of atropine-like side effects at thetherapeutic doses.

(e) Toxicological data of this compound are as follows: LD intravenouslyis 10 mg./ kg. in cat (400 times the minimal effective dose), 12.1mg./kg. in mouse and 11.6 mg./kg. in rat. In the case of thesubcutaneous and oral route, LD in mouse appeared to be higher than 400mg./kg. Death seems to be due to respiratory muscle paralysis; heartactivity is the last that disappears.

A first method for producing the new compounds of this inventioncomprises reacting a p-phenylphenacyl halide of the formula:

wherein X is a halogen, with a tropyl tropate in the presence of aninert solvent at a temperature in the range of from about 0 C. to 60 C.

In accordance with a second method, the compounds of the presentinvention may be prepared by reacting the tropine with ap-phenylphenacyl halide, condensing the Example [I 5.50 g. (0.02 mole)of p-phenylphenacyl bromide were dissolved in 56 cc. of anhydrousacetone previously heated to about 40 C. This solution was added, withstirring, to a solution of 5.70 g. (0.02 mole) of l-hyoscyamine in 43cc. of anhydrous acetone; the reaction solution was maintained at 45 C.and stirred for about six hours.

After standing overnight in the refrigerator, the precipitate wascollected by filtration and dried in vacue at 60 C. Yield: 10.2 g.;M.P.=193 -l94 C. The specific rotation determined in a solution of DMF(dimethylformamide) containing 500 mg. of the product in 10 ml. is 4.7(i0.3) at 25 C.

The compounds of the present invention are compatible with othertherapeutic agents which may be used in connection therewith, such asanalgesics, sedatives, tranquillizing agents, antacids and bulkyconstipants, and may be administered orally, subcutaneously,intravenously or rectally in any of the known pharmaceutical formsgenerally employed for these modes of administration.

In accordance with the present invention the tropyl tropate derivativesmay be associated with a carrier which may be either a solid material ora sterile parenteral liquid. The compositions may be in the form oftablets, capsules, suppositories, vials or other dosage forms. Liquiddiluents,

such as sterile water, are employed in sterile condition for 7 Example111 Aqueous solutions for parenteral use may have the followingcompositions:

pPheny1phenacyl-l-hyoscyarninium bromide mg 2 Distilled water q.s. ad ml2 The vials were sterilized for 20 minutes at 120 C.

p-Phenylphenacyl-l-hyoscyaminium bromide mg 10 Distilled water, q.s. adml 10 The solution was perfectly stable over the period of observation(4 and 9 months at +45 and +55 C.; 15 months at +37 C.).

(C) Since product (K) is very slightly soluble in water (0.18%), it isnecessary to dissolve it in mixtures of organic solvents and water inorder to obtain more concentrated solutions:

p-Phenylphenacyl-l-hyoscyaminium bromide m 10 NaCl mg 14 Propyleneglycol, water for injection, 50% v./v.

q.s. ad ml 2 (D) An aqueous solution for parenteraluse and containing ananalgesic had the following composition:

p-Phenylphenacyl-l-hyoscyaminium bromide mg 2 l-phenyl-2,3dimethyl--pyrazolene-4methylaminomethanesulfonate sodium (analgesic) mg1 Sterile water q.s. ad ml.. 2

The vials were sterilized for 1 hour at 100 C.

Example IV Tablets comprising the compounds of the present invention,suitable for oral administration, may have the following compositions:

Mg. p-Phenylphenacyl-l-hyoscyarninium bromide 2 Kaolin 83 Starch 15 Talc1O Magnesium stearate The powders were mixed, granulated and tableted bywetgranulation.

Mg. p-Phenylphenacyl-l-hyoscyaminium bromide 5 Lactose 54 Starch 34Cielatin 3 Talc 3 Mg stearate 1 This preparation, made bywet-granulation, showed no deterioration when aged for 6 and 12 monthsat +37 C. and C.

Example V Compositions suitable for rectal use had the followingcomposition per suppository p-Phenylphenacyl-l-hyoscyaminium bromidemg.. 3 Triesters of glycerol and fatty acids q.s. ad g 2.5

wherein X is a halogen.

2. The compound of claim 1 wherein the halogen is bromine.

3. p-Phenylphenacyl-hyoscyaminium bromide. 4.p-Phenylphenacyl-dl-tropyltropinium bromide.

References Cited UNITED STATES PATENTS 2,903,394 9/1959 Johnston et al.260-292 WALTER A. MODANCE, Primary Examiner.

A. L. ROTMAN, Assistant Examiner.

1. A COMPOUND OF THE FORMULA: